title: HPLC method for determination of lipoxygenase positional specific products.
abstract: Mammalian lipoxygenases (LOXs) play an important role in physiological and pathological processes through the biosynthesis of lipid mediators-leukotrienes, lipoxins and other arachidonic acid derivatives.There are four major families of LOXs that can be analyzed through the production of hydroxyeicosatetraenoic acids (HETEs). No analytical method to detect 5-, 8-, 12- and 15-HETE in one run has been published to date. The HPLC method combines reversed-phase separative column Nucleosil 120-5 C18 and NP column Zorbax Rx.SIL for identification. This conjunction enables separation of 12-HETE and 15-HETE to the baseline which is essential in 12/15-LOX research and elution of all four HETEs in one run. The method was successfully tested on partially purified LOXs from rat lung cytosol.
authors.affiliation: Faculty of Pharmacy, Department of Cellular and Molecular Biology of Drugs, Comenius University in Bratislava, Bratislava, Slovak Republic. hoffman@fpharm.uniba.sk
authors.forename: Peter
authors.initials: P
authors.lastname: Hoffman
authors.forename: Drahomíra
authors.initials: D
authors.lastname: Rauová
authors.forename: Lýdia
authors.initials: L
authors.lastname: Bezáková
authors.forename: Marek
authors.initials: M
authors.lastname: Obložinský
authors.forename: Peter
authors.initials: P
authors.lastname: Mikuš
mesh_terms: D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid / Q000737:chemistry*; D000818:Animals; D016718:Arachidonic Acid / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D056148:Chromatography, Reverse-Phase / Q000379:methods; D006893:Hydroxyeicosatetraenoic Acids / Q000737:chemistry*; D008084:Lipoxygenase / Q000737:chemistry* / Q000378:metabolism*; D008168:Lung / Q000201:enzymology / Q000378:metabolism; D008297:Male; D051381:Rats; D017208:Rats, Wistar
chemical_list: D006893:Hydroxyeicosatetraenoic Acids; D016718:Arachidonic Acid; D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; C025984:15-hydroxy-5,8,11,13-eicosatetraenoic acid; D008084:Lipoxygenase
keywords: Hydroxyeicosatetraenoic acid; Rat lung; Reversed-phase; Straight-phase; Tissue